Optimization of Formaldehyde Cross-Linking for Protein Interaction Analysis of Non-Tagged Integrin 1
Figure 5
Analysis of integrin complexes (CL) from Jurkat cells and human platelets. (a) Jurkat cells were treated with or without 0.4% formaldehyde (PFA) before lysis. Immunoprecipitation was performed using 2 mg lysate and 5 g of JB1B and analyzed by SDS PAGE. The indicated area was excised and analyzed by mass spectrometry. (b) Anti-integrin immunoblot (JB1A, anti-mouse Alexa Fluor 680) of JB1B pull-downs performed on human platelets treated with and without 0.4% formaldehyde (PFA). 100 g platelet lysate and 0.85 mg JB1B were used. Complexes containing integrin are indicated by the arrow. (c) SDS PAGE analysis of JB1B immunoprecipitates from noncross-linked and cross-linked human platelets. Immunoprecipitation was performed using 0.7 mg lysate and 4.25 g JB1B. The indicated band was excised and examined with mass spectrometry.
